RESUMEN
The oral bioavailability of berberine is quite low due to extensive first-pass metabolism. To increase the bioavailability of berberine (BBR), the efficacy of rectal administration that can avoid intestinal and hepatic first-pass metabolism partly was evaluated using BBR sulfate in rats. BBR sulfate was administered intravenously (1 mg/kg as BBR), orally (10 mg/kg as BBR) and rectally (1, 3, or 10 mg/kg as BBR) using Witepsol® H15 suppository base to evaluate bioavailability in rats. Concentrations of BBR in plasma were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). When BBR sulfate was administered orally, the average oral bioavailability was 0.26%. When BBR sulfate was administered rectally, the average bioavailabilities were 17.0% at 1 mg/kg, 24.3% at 3 mg/kg, and 12.3% at 10 mg/kg as BBR, respectively. Thus, rectal administration of BBR sulfate greatly increased the bioavailability of BBR as compared with oral administration, which would also increase the pharmacological activities of BBR in vivo.
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Berberina , Ratas , Animales , Ratas Sprague-Dawley , Cromatografía Liquida , Disponibilidad Biológica , Administración Rectal , Espectrometría de Masas en Tándem/métodos , Administración Oral , SulfatosRESUMEN
Objective To assess whether neurological maturation and development are accelerated in fetal growth restriction (FGR) in utero using four-dimensional (4D) ultrasound. Methods The facial expressions of 50 appropriate-for-gestational-age (AGA) and 34 FGR fetuses aged between 28 and 35 gestational weeks were assessed using 4D ultrasound. Subsequently, they were differentiated into two gestational age groups (19 aged 28-31 weeks and 31 aged 32-35 weeks in AGA, and 15 aged 28-31 weeks and 19 aged 32-35 weeks in FGR). Fifteen-minute recordings were made, and the frequencies of seven facial expressions during that time were examined. Correlation analysis was conducted to assess the changing number of significant correlations with advancing gestation in each group. Results Out of 21 combinations of facial expressions at 28-31 and 32-35 gestational weeks in AGA fetuses, there was only three significant correlations. Similarly, only one was noted out of 21 combinations at 28-31 gestational weeks, but there were seven significant correlations in FGR fetuses at 32-35 weeks (P=0.018). However, there was no significant difference in the number of correlations of facial expressions between AGA (3/21) and FGR (7/21) fetuses at 32-35 weeks of gestation (P=0.147). Conclusion Our results suggest that the number of significant correlations of fetal facial expressions in FGR fetuses increases more compared with that in AGA fetuses at 32-35 weeks of gestation. Due to the acceleration of neurological maturation and development in FGR fetuses, the control of facial expressions by the brain may be more evident compared with AGA fetuses at 32-35 weeks of gestation. However, the data and their interpretation in the present study should be taken with some degree of caution because of the small number of subjects studied. Further studies involving a larger sample size are needed to obtain strong or additional evidence.
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Expresión Facial , Desarrollo Fetal , Retardo del Crecimiento Fetal/diagnóstico por imagen , Sistema Nervioso/embriología , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Imagenología Tridimensional , Embarazo , Ultrasonografía Prenatal , Adulto JovenRESUMEN
Objective To assess fetal behavioral changes in response to vibroacoustic stimulation (VAS) in normal singleton pregnancies using four-dimensional (4D) ultrasound. Methods Ten types of fetal movements and facial expressions in 68 healthy pregnant women between 24 and 40 weeks were studied using 4D ultrasound for 3 min before and after 3-s VAS. The frequencies of mouthing, yawning, tongue expulsion, back arch, jerky arm movement, startle movement, smiling, scowling, hand-to-face movement, and blinking were evaluated. The fetuses were subdivided into four gestational age groups (24-27, 28-31, 32-35, and ≥36 weeks). Comparison of the frequencies of the fetal behaviors before and after the stimulation in each gestational age group was conducted to detect the response to stimulation with advancing gestation. Results There were no significant differences in the frequency of each fetal behavior before and after VAS at 24-27, 28-31, and 32-35 weeks of gestation. However, the frequencies of blinking and startle movements were significantly higher after VAS in the 36-40 gestational age group (P < 0.05). Conclusion The age of 36 weeks of gestation might represent an advanced stage of brain and central nervous system development and maturation as the response to stimuli is prominent at this age compared with earlier gestation.
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Movimiento Fetal , Ultrasonografía Prenatal , Femenino , Edad Gestacional , Humanos , EmbarazoRESUMEN
Objective To assess the characteristics of twin fetal facial expressions at 30-33 + 6 weeks of gestation using four-dimensional (4D) ultrasound to clarify twin fetal brain development and maturation. Methods Frequencies of seven fetal facial expressions were studied using 4D ultrasound for 15 minutes in 30 singleton pregnancies and 18 twin pregnancies [four monochorionic diamniotic (MD) and 14 dichorionic diamniotic (DD) twins] at 30-33 + 6 weeks of gestation. Comparison of the frequency in each facial expression was performed between singleton and twin fetuses. Results Mouthing was the most frequent facial expression at 30-33 + 6 weeks of gestation, followed by blinking in twin and singleton fetuses. Both facial expressions were significantly more frequent than other expressions (P < 0.05). The frequencies of mouthing and scowling in twin fetuses were significantly lower than those in singleton fetuses, but there were no significant differences in the frequencies of the five other facial expressions between the fetal groups. Conclusion Our results suggest that restricted twin fetal behavior before 20 weeks of gestation may still affect the frequencies of twin facial expressions early in the third trimester of pregnancy. Moreover, the frequencies of facial expressions in twins are different from those of singleton fetuses.
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Encéfalo/embriología , Expresión Facial , Desarrollo Fetal , Tercer Trimestre del Embarazo , Embarazo Gemelar , Adulto , Femenino , Movimiento Fetal , Madurez de los Órganos Fetales , Humanos , Masculino , Embarazo , Ultrasonografía Prenatal/métodosRESUMEN
AIM: To assess the effect of psychotropic drugs on fetal behavior using four-dimensional (4D) ultrasound in the third trimester of pregnancy. METHODS: Fetal behavior was assessed using Kurjak's antenatal neurodevelopmental test (KANET) using 4D ultrasound between 28 and 36 weeks of gestation. Thirty healthy (control group) and 10 psychotropic-drug-administered pregnant (case group) women were studied. The total value of the KANET score and values of each parameter (eight parameters) were compared between the two groups. RESULTS: The total KANET score was normal (except for one fetus in the case group: total score of 9) in both groups, and there was no significant difference in the total KANET score. When individual KANET parameters were compared, no significant differences were noted in any of the eight parameters. CONCLUSION: Our results showed that there is no difference in fetal behavior between fetuses of normal pregnant women and those of psychotropic-drug-administered pregnant women in the third trimester of pregnancy. These results suggest that psychotropic drugs may not affect fetal behavioral development in utero. However, the data and their interpretation in the present study should be taken with some degree of caution because of the small number of subjects studied. Further studies involving a larger sample size are needed to assess the effect of psychotropic drugs on fetal neurobehavior during pregnancy.
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Desarrollo Fetal/efectos de los fármacos , Movimiento Fetal/efectos de los fármacos , Psicotrópicos , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Japón , Evaluación de Resultado en la Atención de Salud , Embarazo , Tercer Trimestre del Embarazo , Embarazo de Alto Riesgo , Atención Prenatal/métodos , Psicotrópicos/administración & dosificación , Psicotrópicos/efectos adversos , Psicotrópicos/clasificación , Proyectos de Investigación , Ultrasonografía Prenatal/métodosRESUMEN
The hexactinellids are a diverse group of predominantly deep sea sponges that synthesize elaborate fibrous skeletal systems of amorphous hydrated silica. As a representative example, members of the genus Euplectella have proved to be useful model systems for investigating structure-function relationships in these hierarchically ordered siliceous network-like composites. Despite recent advances in understanding the mechanistic origins of damage tolerance in these complex skeletal systems, the details of their synthesis have remained largely unexplored. Here, we describe a previously unidentified protein, named "glassin," the main constituent in the water-soluble fraction of the demineralized skeletal elements of Euplectella. When combined with silicic acid solutions, glassin rapidly accelerates silica polycondensation over a pH range of 6-8. Glassin is characterized by high histidine content, and cDNA sequence analysis reveals that glassin shares no significant similarity with any other known proteins. The deduced amino acid sequence reveals that glassin consists of two similar histidine-rich domains and a connecting domain. Each of the histidine-rich domains is composed of three segments: an amino-terminal histidine and aspartic acid-rich sequence, a proline-rich sequence in the middle, and a histidine and threonine-rich sequence at the carboxyl terminus. Histidine always forms HX or HHX repeats, in which most of X positions are occupied by glycine, aspartic acid, or threonine. Recombinant glassin reproduces the silica precipitation activity observed in the native proteins. The highly modular composition of glassin, composed of imidazole, acidic, and hydroxyl residues, favors silica polycondensation and provides insights into the molecular mechanisms of skeletal formation in hexactinellid sponges.
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Histidina/química , Poríferos/química , Proteínas/química , Dióxido de Silicio/química , Secuencia de Aminoácidos , Aminoácidos/química , Animales , Ácido Aspártico/química , Sitios de Unión , Clonación Molecular , ADN Complementario/química , Electroforesis en Gel de Poliacrilamida , Epítopos/química , Geografía , Concentración de Iones de Hidrógeno , Hidrólisis , Datos de Secuencia Molecular , Péptidos/química , Prolina/química , Procesamiento Proteico-Postraduccional , Proteínas/genética , Proteínas Recombinantes/química , Homología de Secuencia de Aminoácido , Solubilidad , Temperatura , Treonina/químicaRESUMEN
Objective To evaluate fetal behavioral differences between singleton and twin fetuses before 20 weeks of gestation using four-dimensional (4D) ultrasound. Methods 4D ultrasound was used to examine fetal movements in 58 singleton and 48 twin normal fetuses at 12-19 weeks. The frequencies of eight fetal movements were assessed through 15-min recordings. The fetuses were divided into two gestational age groups (12-13 and 14-19 weeks) to evaluate the changes with advancing gestation in twin versus singleton fetuses. Results Arm and general movements were the most frequent movements in singleton fetuses, whereas only general movement was significantly more frequent than the other seven fetal movements in twin fetuses at 12-13 weeks. At 14-19 weeks, frequencies of arm and leg movements were significantly higher than those of the other six movements in singleton fetuses, while only arm movement was significantly more frequent than the other fetal movements in twin fetuses. Comparisons of fetal movements between singleton and twin fetuses revealed that only arm movement showed a significant difference at 12-13 weeks, while the frequencies of all movements in singleton fetuses were significantly higher than those in twin fetuses at 14-19 weeks. Conclusion Our results suggest that the limitation of available space and crowding of twin fetuses with advancing gestation may have a marked impact on twin fetal movements compared with singleton fetuses, even in the first half of pregnancy. Further studies are needed to assess whether decreased fetal movements in twin pregnancy can affect fetal and neonatal development and maturation before and after birth.
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Movimiento Fetal , Embarazo Gemelar/fisiología , Femenino , Humanos , Embarazo , Ultrasonografía PrenatalRESUMEN
Objective To assess the usefulness of the antenatal fetal neurodevelopmental test for the prediction of postnatal developmental disabilities. Methods Fetal behavior was assessed with Kurjak's antenatal neurodevelopmental test (KANET) using four-dimensional ultrasound between 28 and 38 weeks of gestation. A score range of 0-5 was characterized as abnormal, from 6 to 9 was considered borderline, and 10-16 was normal. After birth, follow-up was conducted for at least 2 years in all fetuses. Results There were 337 normal (95.47%) and 16 borderline (4.53%) cases among the 353 cases studied, whereas there was no abnormal case. Five cases with postnatal developmental disabilities (one case of Werdig-Hoffmann disease diagnosed just after delivery, one case of autism spectrum disorder diagnosed at 24 months, one case of Ullrich congenital muscular dystrophy diagnosed at 9 months and two cases of developmental disorders diagnosed at age 3 and 18 months) were noted among the 337 normal cases (1.48%), whereas three cases with developmental disabilities (one case of motor development delay diagnosed at 6 months, one case of Duchenne muscular dystrophy diagnosed at 18 months and one case of autism spectrum disorder diagnosed at age 30 months) were found among the 16 borderline cases (18.75%). There was a significant difference in the prevalence of postnatal developmental disabilities between the normal and borderline KANET groups (P<0.001). Conclusion Our results suggest that the KANET assessment may be a useful diagnostic modality for the prediction of postnatal developmental disabilities.
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Trastorno del Espectro Autista , Discapacidades del Desarrollo , Desarrollo Fetal , Movimiento Fetal , Feto , Sistema Nervioso/crecimiento & desarrollo , Ultrasonografía Prenatal/métodos , Adulto , Trastorno del Espectro Autista/diagnóstico , Trastorno del Espectro Autista/epidemiología , Preescolar , Discapacidades del Desarrollo/clasificación , Discapacidades del Desarrollo/diagnóstico , Discapacidades del Desarrollo/epidemiología , Femenino , Feto/diagnóstico por imagen , Feto/fisiología , Edad Gestacional , Humanos , Lactante , Japón/epidemiología , Masculino , Valor Predictivo de las Pruebas , Embarazo , Diagnóstico Prenatal/métodos , Prevalencia , PronósticoRESUMEN
We describe here a novel chemotype with substituted benzimidazole scaffold for nonsteroidal farnesoid X receptor (FXR) antagonists starting from the identification of a screening hit, BB-4. Structure diversity in four regions A-D of BB-4 or 1 is discussed. In particular, regions A and C had an effect on an antagonism against FXR as demonstrated by the derivatives represented by 7 and 15, respectively. Thus, compound 19 arising from the combination of regions A and C underscored an important fact on antagonism against FXR, also showing the reduced small heterodimer partner and the increased cholesterol 7α-hydroxylase expression levels.
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Bencimidazoles/farmacología , Descubrimiento de Drogas , Receptores Citoplasmáticos y Nucleares/antagonistas & inhibidores , Bencimidazoles/química , Línea Celular Tumoral , Colesterol 7-alfa-Hidroxilasa/genética , Colesterol 7-alfa-Hidroxilasa/metabolismo , Transferencia Resonante de Energía de Fluorescencia , Humanos , Espectroscopía de Protones por Resonancia Magnética , ARN Mensajero/genética , Relación Estructura-ActividadRESUMEN
We present our experience of using the HDLive silhouette inversion mode to assess complete hydatidiform mole early in pregnancy. The HDLive silhouette inversion mode clearly depicted the number, size, and spatial position of molar vesicles, compared with conventional two-dimensional sonography or the HDLive inversion mode. Moreover, spatial relationships among molar vesicles, intrauterine anechoic fluid collection, and the uterine wall enabled the clear localization of the lesion. This technique provides new insights, and has the potential to supplement conventional two-dimensional sonography in the diagnosis of complete hydatidiform mole.
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Mola Hidatiforme/diagnóstico por imagen , Imagenología Tridimensional/métodos , Ultrasonografía Prenatal/métodos , Neoplasias Uterinas/diagnóstico por imagen , Adulto , Diagnóstico Diferencial , Femenino , Humanos , Embarazo , Útero/diagnóstico por imagenRESUMEN
AIM: To assess the frequency of fetal movement and reproducibility of fetal movement counting in normal singleton pregnancies early in the second trimester using four-dimensional (4D) ultrasound. METHODS: Twenty-nine singleton pregnancies were studied for 15 min employing 4D ultrasound at 14-16 (19 cases) and 17-19 (10 cases) weeks of gestation. The frequencies of eight fetal movements (head anteflexion, head retroflexion, body rotation, hand to face movement, general movement, isolated arm movement, isolated leg movement and mouthing movement) were evaluated. RESULTS: The most frequent fetal movements were isolated arm movements at 14-16 and 17-19 weeks' gestation. There was a significant difference only in the frequency of mouthing movement between 14-16 and 17-19 weeks' gestation (P<0.05). All fetal movements showed intra- and inter-class correlation coefficients greater than 0.87, with good intra- and inter-observer agreements. CONCLUSION: The difference in the frequency of mouthing movement at 14-16 and 17-19 weeks' gestation may be due to increasing fetal swallowing because of the increasing amniotic fluid early in the second trimester of pregnancy. To the best of our knowledge, this is the first study on the reproducibility assessment of fetal movement counting using 4D ultrasound. However, the data and their interpretation in the present study should be taken with some degree of caution because of the small number of subjects studied. Further studies involving a larger sample size are needed to assess the reproducibility of fetal movement counting using 4D ultrasound.
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Movimiento Fetal , Ultrasonografía Prenatal , Femenino , Humanos , Imagenología Tridimensional , Embarazo , Segundo Trimestre del EmbarazoRESUMEN
We present a case of antenatal diagnosis of idiopathic dilatation of the pulmonary artery by three-dimensional power Doppler imaging (HDliveFlow with silhouette mode). Two-dimensional sonography clearly demonstrated a 10.4-mm-diameter fetal pulmonary artery (PA), whereas the aorta (Ao) diameter was 5.04 mm. HDliveFlow clearly demonstrated the spatial relationships and different sizes of PA and Ao. The PA size returned to normal (8.0 mm) on the 11th day after birth (Ao: 9.0 mm). HDliveFlow may be an adjunctive tool to two-dimensional sonography to diagnose abnormalities of fetal great vessels. © 2016 Wiley Periodicals, Inc. J Clin Ultrasound 45:121-123, 2017.
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Cardiopatías Congénitas/diagnóstico por imagen , Imagenología Tridimensional/métodos , Arteria Pulmonar/anomalías , Ultrasonografía Doppler/métodos , Ultrasonografía Prenatal/métodos , Adulto , Femenino , Humanos , Masculino , Embarazo , Arteria Pulmonar/diagnóstico por imagenRESUMEN
The report is the first of purification, overproduction, and characterization of a unique γ-butyrobetainyl CoA synthetase from soil-isolated Agrobacterium sp. 525a. The primary structure of the enzyme shares 70-95% identity with those of ATP-dependent microbial acyl-CoA synthetases of the Rhizobiaceae family. As distinctive characteristics of the enzyme of this study, ADP was released in the catalytic reaction process, whereas many acyl CoA synthetases are annotated as an AMP-forming enzyme. The apparent Km values for γ-butyrobetaine, CoA, and ATP were, respectively, 0.69, 0.02, and 0.24 mM.
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Acilcoenzima A/metabolismo , Adenosina Difosfato/metabolismo , Agrobacterium/enzimología , Proteínas Bacterianas/metabolismo , Betaína/análogos & derivados , Carnitina/metabolismo , Coenzima A Ligasas/metabolismo , Microbiología del Suelo , Acilcoenzima A/química , Adenosina Difosfato/química , Agrobacterium/genética , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Proteínas Bacterianas/aislamiento & purificación , Betaína/química , Betaína/metabolismo , Carnitina/química , Clonación Molecular , Coenzima A Ligasas/genética , Coenzima A Ligasas/aislamiento & purificación , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Concentración de Iones de Hidrógeno , Cinética , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Especificidad por SustratoRESUMEN
From investigation of 60 filamentous fungi, we identified Fusarium merismoides var. acetilereum, which uses 4-N-trimethylamino-1-butanol (TMA-butanol) as the sole source of carbon and nitrogen. The fungus produced NAD(+)-dependent TMA-butanol dehydrogenase (DH) when it was cultivated in medium containing TMA-butanol. The enzyme showed molecular mass of 40 kDa by SDS-PAGE and 160 kDa by gel filtration, suggesting that it is a homotetramer. TMA-butanol DH is stable at pH 7.5-9.0. It exhibits moderate stability with respect to temperature (up to 30 °C). Additionally, it has optimum activity at 45 °C and at pH 9.5. The enzyme has broad specificity to various alkyl alcohols and amino alkyl alcohols, and the carbon chains of which are longer than butanol. Moreover, the activity is strongly inhibited by oxidizing agents, carbonyl and thiol modulators, and chelating agents. This report is the first study examining TMA-butanol DH from eukaryotic microbes.
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Oxidorreductasas de Alcohol/química , Oxidorreductasas de Alcohol/aislamiento & purificación , Fusarium/enzimología , Oxidorreductasas de Alcohol/genética , Amino Alcoholes/química , Carbono/química , Fusarium/química , Concentración de Iones de Hidrógeno , Cinética , Especificidad por Sustrato , TemperaturaRESUMEN
A ß-peptidyl aminopeptidase, a peptidase belonging to the P1 family, catalyzes aminolysis in accordance with its hydrolytic activity. We specifically examined ß-peptidyl aminopeptidase of Pseudomonas aeruginosa PAO1 (BapF) to assess the effects of mutation of catalytic Ser with Cys or Thr on its catalytic ability. Recombinant BapF and its S237C mutant exhibited p-nitroaniline release activity toward ß-homo-Gly-p-nitroanilide (ßhGly-pNA), but the products of the enzyme reaction differed completely from one another. Wild-type BapF showed ßhGly-ßhGly-pNA synthetic activity, but the product vanished in a few minutes and converted to free ßhGly. In contrast, the product ßhGly-ßhGly-pNA was synthesized by S237C BapF efficiently without degradation, indicating that because of the mutation, the enzyme came to recognize only the amine group as an acyl acceptor instead of water. Furthermore, a difference in acyl acceptor preference between that of wild type and S237C BapF was observed. When using cysteamine as an acyl acceptor, ßhGly-cysteamine was synthesized only in the reaction using S237C BapF. In contrast, S237C BapF was unable to synthesize ßhGly-cystamine when using cystamine as an acyl acceptor, although it was synthesized by wild-type BapF. Such a dynamic change in the acyl acceptor by the mutation of catalytic Ser with Cys is regarded as a unique feature of family P1 peptidases.
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Aminopeptidasas/química , Aminopeptidasas/metabolismo , Cisteína/química , Pseudomonas aeruginosa/enzimología , Aminopeptidasas/genética , Catálisis , Dominio Catalítico , Mutación , SerinaRESUMEN
Pleurotus eryngii serine aminopeptidase that has peptide bond formation activity, redesignated as eryngase, was cloned and expressed. Eryngase has a family S9 peptidase unit in the C-terminal region having a catalytic triad of Ser, Asp, and His. In the phylogenetic relations among the subfamilies of family S9 peptidase (S9A, prolyl oligopeptidase; S9B, dipeptidyl peptidase; S9C, acylaminoacyl peptidase; S9D, glutamyl endopeptidase), eryngase existed alone in the neighbor of S9C subfamily. Mutation of the active site Ser524 of the eryngase with Ala eliminated its catalytic activity. In contrast, S524C mutant maintained low catalytic activity. Investigation of aminolysis activity using l-Phe-NH2 as a substrate showed that S524C mutant exhibited no hydrolysis reaction but synthesized a small amount of l-Phe-l-Phe-NH2 by the catalysis of aminolysis. In contrast, wild-type eryngase hydrolyzed the product of aminolysis l-Phe-l-Phe-NH2. Results show that the S524C mutant preferentially catalyzed aminolysis when on an l-Phe-NH2 substrate.
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Aminopeptidasas/genética , Aminopeptidasas/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Péptido Hidrolasas/genética , Péptido Hidrolasas/metabolismo , Pleurotus/enzimología , Pleurotus/genética , Secuencia de Aminoácidos , Aminopeptidasas/química , Secuencia de Bases , Dominio Catalítico/genética , Cromatografía Líquida de Alta Presión , Clonación Molecular , Proteínas Fúngicas/química , Cromatografía de Gases y Espectrometría de Masas , Datos de Secuencia Molecular , Mutación , Péptido Hidrolasas/química , Pleurotus/clasificación , Alineación de Secuencia , Serina/química , Serina/genéticaRESUMEN
L-Carnitine dehydrogenase (CDH) is as an excellent tool for L-carnitine (L-Car) estimation. To date, four CDHs have been identified, that share 45 % homology of their proteins. Here 42 conserved residues of CDH from Xanthomonas translucens (Xt-CDH) were substituted successively with alanine. The resultant mutants were analyzed for catalytic activity. Active mutants were evaluated for their influence on L-Car affinity. Twenty-three mutants with reduced affinity toward L-Car were subjected to detailed kinetic analysis. Analytical data implied that all mutants had increased K m values. The mutants of R193A, E196A, W199A, R200A, F249A, and F253A that produced the greatest L-Car affinity disruption (K m > 200-folds of Xt-CDH) clustered near the putative active site. This information can provide a solid basis for the rational design of mutagenic investigation to improve CDHs.
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Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Carnitina/metabolismo , Análisis Mutacional de ADN , Xanthomonas/enzimología , Sustitución de Aminoácidos , Dominio Catalítico , Cinética , Xanthomonas/genéticaRESUMEN
Upshaw-Schulman syndrome (USS) involves a congenital deficiency of von Willebrand factor-cleaving metalloprotease (ADAMTS13) activity due to gene mutations. Female patients develop overt thrombotic thrombocytopenic purpura (TTP) caused by a decline of ADAMTS13 activity in pregnancy. A 23-year-old nulliparous Japanese woman died due to severe, rapid progression of TTP with intrauterine fetal death at 20 weeks of gestation after its onset, even though she underwent intensive treatment which included plasma exchange. She had a history of idiopathic thrombocytopenic purpura at the age of 3 years. The patient's ADAMTS13 activity was of very low level. It should be borne in mind that there is the possibility of rapidly progressive fulminant USS during pregnancy.
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Muerte Fetal/etiología , Complicaciones Hematológicas del Embarazo/fisiopatología , Púrpura Trombocitopénica Trombótica/fisiopatología , Proteínas ADAM/sangre , Proteínas ADAM/genética , Proteína ADAMTS13 , Adulto , Terapia Combinada , Progresión de la Enfermedad , Resultado Fatal , Femenino , Humanos , Mutación , Intercambio Plasmático , Embarazo , Complicaciones Hematológicas del Embarazo/sangre , Complicaciones Hematológicas del Embarazo/genética , Complicaciones Hematológicas del Embarazo/terapia , Segundo Trimestre del Embarazo , Púrpura Trombocitopénica Trombótica/sangre , Púrpura Trombocitopénica Trombótica/genética , Púrpura Trombocitopénica Trombótica/terapia , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
BACKGROUND: Our objective is to present our first experience of adnexal tumors reconstructed employing the HDlive rendering mode. METHODS: Seven adnexal tumors (one case each of chocolate cyst, benign cystic teratoma, serous cystadenofibroma, and torsion of the hydrosalpinx, and three cases of ovarian cancer) were studied using the HDlive rendering mode. RESULTS: In the case of the chocolate cyst, the smooth inner wall with numerous tiny projections was clearly identified. In the case of benign cystic teratoma, a smooth hair ball inside the cyst was evident. In the case of the serous cystadenofibroma, a smooth, solid projection and relatively smooth septum were noted. In the three cases of ovarian cancer, numerous irregular papillae and irregular, thick septae were depicted. In the case of the bilateral hydrosalpinx with torsion of the right tube, different fluid contents were recognized. CONCLUSION: The HDlive rendering mode provides anatomically realistic features such as macroscopic specimens for physicians and pathologists due to the complete depiction of the inner surface of the adnexal tumors.
RESUMEN
N-Acyl-D-amino acid amidohydrolases (D-aminoacylases) are often used as tools for the optical resolution of D-amino acids, which are important products with applications in industries related to medicine and cosmetics. For this study, genes encoding D-aminoacylase were cloned from the genomes of Streptomyces spp. using sequence-based screening. They were expressed by Escherichia coli and Streptomyces lividans. Almost all of the cell-free extracts exhibit hydrolytic activity toward N-acetyl-(Ac-)D-Phe (0.05-6.32 µmol min(-1) mg(-1)) under conditions without CoCl2. Addition of 1 mM CoCl2 enhanced their activity. Among them, the highest activity was observed from cell-free extracts prepared from S. lividans that possess the D-aminoacylase gene of Streptomyces sp. 64E6 (specific activities were, respectively, 7.34 and 9.31 µmol min(-1) mg(-1) for N-Ac-D-Phe and N-Ac-D-Met hydrolysis). Furthermore, when using glycerol as a carbon source for cultivation, the recombinant enzyme from Streptomyces sp. 64E6 was produced in 4.2-fold greater quantities by S. lividans than when using glucose. D-Aminoacylase from Streptomyces sp. 64E6 showed optimum at pH 8.0-9.0. It was stable at pH 5.5-9.0 up to 30 °C. The enzyme hydrolyzed various N-acetyl-D-amino acids that have hydrophobic side chains. In addition, the activity toward N-chloroacetyl-D-Phe was 2.1-fold higher than that toward N-Ac-D-Phe, indicating that the structure of N-acylated portion of substrate altered the activity.